Department of Surgery, Tongji Hospital, Tongji University School of Medicine, Tongji University, Shanghai 200065, P.R. China.
Oncol Rep. 2019 Oct;42(4):1475-1486. doi: 10.3892/or.2019.7249. Epub 2019 Jul 25.
Gallbladder cancer (GBC) is a lethal aggressive malignant neoplasm of the biliary tract. Potential prognostic markers and therapeutic targets for this disease are urgently required. Cancer‑associated fibroblasts (CAFs) play a key role in tumorigenesis and the development of cancer. Nicotinamide adenine dinucleotide phosphate oxidase 1 (NOX1) expression has been reported to be involved in tumorigenesis and useful for tumor prognosis. However, NOX1 expression in the stroma of GBCs, particularly gallbladder cancer‑associated fibroblasts (GCAFs), and its prognostic significance in GBC patients remains unclear. In the present study, NOX1 expression in the stroma of human gallbladder lesions in vivo was investigated, as well as in GCAFs and co‑cultures of GBC‑SD+GCAFs in vitro, and their correlation with clinicopathological parameters and the prognosis of GBC patients were evaluated. The results revealed that NOX1 expression was significantly upregulated in the stroma of GBCs compared with precancerous and benign lesions of the gallbladder; NOX1 expression was localized to gallbladder stromal fibroblasts expressing α‑smooth muscle actin and fibroblast secreted protein‑1. Furthermore, these observations were confirmed by the fact that NOX1 expression was upregulated in GCAFs as determined by Affymetrix gene profile chip analysis and reverse transcription‑quantitative PCR. In addition, overexpression was observed in formed spheroids of GBC‑SD+GCAF co‑cultures by immunohistochemistry and western blotting in vitro. Thus, it was verified that NOX1 expression was upregulated in GCAFs. Furthermore, upregulated stromal NOX1 expression was correlated with aggressive characteristics such as differentiation degree (P=0.042), venous invasion (P=0.041), resection methods (P=0.002), and a lower survival rate (P=0.025, log‑rank test) of patients with GBC. Stromal NOX1 expression (P=0.047) was an independent prognostic factor for the overall survival rate of patients with GBC. GBC patients with upregulated NOX1 expression in GCAFs had a poorer prognosis. These results revealed that stromal NOX1 may be a novel biomarker and/or target, and may contribute to the discovery of new tumor markers and potential targeted therapeutics for human GBCs.
胆囊癌（GBC）是一种致命的侵袭性胆道恶性肿瘤。目前迫切需要针对这种疾病的潜在预后标志物和治疗靶点。癌症相关成纤维细胞（CAFs）在肿瘤发生和癌症发展中起关键作用。烟酰胺腺嘌呤二核苷酸磷酸氧化酶 1（NOX1）的表达已被报道与肿瘤发生有关，并可用于肿瘤预后。然而，GBC 肿瘤基质中（尤其是胆囊癌相关成纤维细胞，GCAFs）NOX1 的表达及其在 GBC 患者中的预后意义尚不清楚。在本研究中，我们研究了体内人胆囊病变基质中 NOX1 的表达，以及体外 GBC-SD+GCAFs 共培养物中的表达，并评估了其与 GBC 患者临床病理参数和预后的相关性。结果显示，与胆囊癌前病变和良性病变相比，GBC 肿瘤基质中 NOX1 的表达显著上调；NOX1 表达定位于表达α-平滑肌肌动蛋白和纤维母细胞分泌蛋白-1 的胆囊基质成纤维细胞。此外，通过 Affymetrix 基因芯片分析和逆转录定量 PCR 证实了 GCAFs 中 NOX1 表达上调的这一观察结果。此外，在体外共培养的 GBC-SD+GCAF 形成的球体中通过免疫组织化学和 Western blot 观察到过表达。因此，证实了 GCAFs 中 NOX1 表达上调。此外，基质中 NOX1 的表达上调与肿瘤侵袭性特征相关，如分化程度（P=0.042）、静脉侵犯（P=0.041）、切除方法（P=0.002）和 GBC 患者的生存率降低（P=0.025，对数秩检验）相关。基质中 NOX1 的表达（P=0.047）是 GBC 患者总生存率的独立预后因素。GBC 患者中 GCAFs 中 NOX1 表达上调与预后不良相关。这些结果表明，基质中的 NOX1 可能是一种新的生物标志物和/或靶点，有助于发现人类 GBC 的新肿瘤标志物和潜在的靶向治疗方法。
